Construction of eukaryotic p75 expression vector and its expression in human PC-3 cells

Objective:To construct recombinant eukaryotic expression vector pcDNA3.1(+)-p75 and express it in prostate cancer (PC-3) cells. Methods: Human p75 cDNA was obtained from the human brain glial cell tumor by RT-PCR and ligated with pcDNA3.1(+). The recombinant eukaryotic expression vector pcDNA3.1(+)-p75 was transferred into PC-3 cells after selection with restriction enzyme digestion and sequencing. The positive clones were isolated with G418. The expression of p75 was verified by immunohistochemical technique and RT-PCR assay. Results: The human p75 cDNA was verified by sequence. Immunohistochemical technique and RT-PCR assay indicated the transcription of p75 gene and the recombinant p75 protein was detected in the transfected PC-3 cells.Conclusion:Recombinant eukaryotic expression vector pcDNA3.1(+)/-p75 is successfully constructed, and the recombined p75 protein is expressed on the surface of PC-3 cells.